Industrial manufacture generates a huge quantity of emulsion wastewater, which causes serious threats to the aquatic ecosystems. Water-in-oil (W/O) and oil-in-water (O/W) emulsions are two major types of emulsions discharged by industries. However, dual separation of W/O and O/W emulsions remains a challenging issue due to the contradictory permselectivity for separating the two emulsions. In the present investigation, the amphiphilicity-derived regional wetting mechanism of water and oil on the amphiphilic collagen fibers was revealed based on the combination of numerous experiments and molecular dynamics (MD) simulations. Electrostatic interactions and van der Waals force were manifested to be the driving forces of regional wetting in the hydrophilic and hydrophobic regions, respectively. The regional wetting endowed amphiphilic collagen fibers with underwater oleophobicity and underoil hydrophilicity, which enabled dual separation of emulsions by selectively retaining the dispersed water phase of W/O emulsions in the hydrophilic regions while the dispersed oil phase of O/W emulsions in the hydrophobic regions. The achieved separation efficiency was higher than 99.98%, and the flux reached 3337.6 L m-2 h-1. Initial wetting status significantly affects the regional wetting-enabled dual separation. Based on the MD simulations, amphiphilic intramolecular conformations of tropocollagen were suggested to be the origins of regional wetting on collagen fibers. Our findings may pave the way for developing high-performance dual separation materials that are promising to be utilized for the practical treatment of emulsion wastewater.